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Visceral leishmaniasis (VL) is an urgent public health concern in Brazil. We evaluated the spatiotemporal distribution of VL to better understand the effects of economic activities related to agriculture, livestock, and deforestation on its incidence in the Brazilian Legal Amazon (BLA). The data on newly confirmed cases of VL in Brazilian municipalities from 2007 to 2020 were extracted from the Brazilian Notifiable Diseases Information System (SINAN) and analyzed. The data on agricultural production (planted area in hectares) and livestock (total number of cattle) were obtained from the Brazilian Institute of Geography and Statistics (IBGE), whereas deforestation data (in hectares) were obtained from the Amazon Deforestation Estimation Project (PRODES). SatScan and the local indicators of spatial association (LISA) were used to identify the spatial and temporal patterns of VL and its relationships with economic and environmental variables. The cumulative incidence rate was found to be 4.5 cases per 100,000 inhabitants. Based on the LISA results, areas with a high incidence of VL and deforestation were identified in the states of Roraima, Pará, and Maranhão. Strengthening deforestation monitoring programs and environmental enforcement actions can help implement public policies to control illegal deforestation and mitigate the socio-environmental vulnerability in the BLA. Therefore, areas identified in this study should be prioritized for controlling VL. [ABSTRACT FROM AUTHOR]

Visceral Leishmaniasis is a serious public health problem caused by Leishmania species parasites. Approximately 500 thousand people get Visceral Leishmaniasis (VL) every year. An effective and reliable vaccine against the disease has still not been formulated. Choosing the right adjuvant is important to increase immunogenicity in vaccines prepared with total antigens. In this study, we investigate the ideal adjuvant for use in vaccine formulations against VL. For this purpose, Leishmania antigens (FTLA) obtained from L. infantum parasites by the freeze-thaw method and three different adjuvants (alum-saponin and calcium phosphate) were used. The effectiveness of the formulations was investigated in vitro by cell viability analysis and determination of nitric oxide and cytokine production abilities in J774 macrophage cells. According to the study results, it was determined that formulations prepared with calcium phosphate produced 72% more NO and approximately 7.2 times more IL-12 cytokine. The results obtained showed that calcium phosphate salts can be used as ideal adjuvants in vaccine research against leishmaniasis. [ABSTRACT FROM AUTHOR]

Introduction: In Ethiopia, visceral leishmaniasis (VL) is a public health concern that has been spreading to new endemic foci in recent years. An estimated 3.2 million people are at risk of infection, with 3700–7400 new cases yearly. Thus, the study aimed to determine the prevalence of VL and associated risk factors among febrile patients attending Metema Hospital, North West Ethiopia. Methods: A hospital-based cross-sectional study was conducted on 404 febrile patients attending Metema Hospital from February 2021 to June 2021. The test for VL was done using an immune-chromatographic test (RK39) according to the manufacturer's instructions (InBios International Inc., USA). An interviewer-administered, pretested questionnaire was used to collect data on risk factors associated with VL. Logistic regression and Chi-square assessed the association between VL and the associated risk factors. Reults: The overall prevalence of visceral leishmaniasis was 18.8% (76/404), with a higher prevalence of VL in males, in the age category between 21 and 30, in study participants who completed elementary school, and in those who earned less than 500 birr monthly compared to their counterparts. Houses with thatched roofs (adjusted odd ratio (AOR) = 17.648, 95CI = 6.549,47.563), houses with mud walls (AOR = 2.538, 95% CI = 1.187–5.411), cattle ownership (AOR = 3.173, 95% CI = 1.286–7.826), dog ownership (AOR = 2,533, 95% CI = 1.256–5.111), presence of Acacia trees near houses (AOR = 1.975, 95% CI:1.004–3.886), presence of Balanites tree (AOR = 3.015, 95% CI = 1.610–5.992), and outdoor sleeping (AOR = 2.259, 95% CI: 1.107–14.607) were the predictors of VL in the present study. Conclusions: In the study area, VL is still very common. Thus, preventing and controlling infection in the area is largely dependent on raising community awareness of VL prevention and control measures and implementing the necessary interventions on the determinants that have been identified. [ABSTRACT FROM AUTHOR]

Purpose: Visceral leishmaniasis (VL) is caused by an intracellular parasite that is transmitted to humans by sandfly bites. It is prevalent throughout Asia, Africa, the Americas, and the Mediterranean area, where 147 million people are at risk of contracting the illness. The manifestation of heterotrophic illness relies on both Leishmania implicated and the host's immunological response, ranging from asymptomatic to severe leishmaniasis with potentially lethal effects. Method: We reviewed the literature (published till 31st December 2023) on the worldwide situation of leishmaniasis, standard and novel detection techniques, and traditional and modern treatment strategies and endeavors to eliminate VL. Moreover, epidemiological data was collected from the World Health Organization's publicly available databases. GraphPad Prism Version 8 was used to analyze and produce figures based on the epidemiological data. Results: Diagnosis of parasites in tissues or serology is commonly employed. Diagnosis by identifying parasite DNA using molecular techniques is becoming more popular. Despite recent findings of L. donovani resistance to pentavalent antimoniate medications, it continues to be the cornerstone in the medical management of VL. Amphotericin B and its lipid formulations, injectable paromomycin, and oral miltefosine are among the new therapy options being researched. The number of reported VL cases has reduced remarkably over the last decade due to human interventions made to eliminate VL. Particularly countries from the South East Asian region have experienced momentous progress in reducing VL cases and eliminating this disease from this region. Owing to the robust elimination programs, countries such as Bangladesh has eliminated VL as a public health concern. India and Nepal are on the verge of its elimination. Conclusion: Rapid diagnosis, effective and inexpensive treatment, simple access to newly discovered medications, appropriate vector control, and a well-designed vaccine are all required for the elimination of this disease burden in impoverished areas of the globe. [ABSTRACT FROM AUTHOR]

Purpose: This case report examines the clinical presentation, diagnosis, treatment, and outcomes of mucocutaneous leishmaniasis with primary oral involvement in HIV-positive and HIV-negative patients diagnosed in Brazil. Methods: We discuss the clinical manifestations, diagnostic methods, and therapeutic strategies, highlighting the clinical and histopathologic diagnostic features and distinct progression patterns based on HIV status. Our findings are compared with patterns observed in other countries, emphasizing the differences between the Americas and Europe, Asia, and Africa. Results: In the Americas, particularly in Brazil, mucocutaneous leishmaniasis often presents with localized oral lesions, even in the presence of systemic immunosuppression, whereas in the Europe, Asia, and Africa, oral involvement is typically associated with visceral leishmaniasis in immunocompromised patients. These differences were due to variations in the parasite species involved. Conclusion: This comparison underscores the importance of regional and immunological factors in diagnosing and managing this neglected infectious disease. [ABSTRACT FROM AUTHOR]

Purpose: Leishmaniasis, caused by the parasite of the genus Leishmania, is a neglected tropical disease which is endemic in more than 60 countries. In South-East Asia, Brazil, and East Africa, it mainly occurs as kala-azar (visceral leishmaniasis, VL), and subsequently as post kala-azar dermal leishmaniasis (PKDL) in a smaller portion of cases. As stated per WHO roadmap, accessibility to accurate diagnostic methods is an essential step to achieve elimination. This study aimed to test the accuracy of a portable minoo device, a small battery-driven, multi-use fluorimeter operating with isothermal technology for molecular diagnosis of VL and PKDL. Methods: Fluorescence data measured by the device within 20 min are reported back to the mobile application (or app) via Bluetooth and onward via the internet to a backend. This allows anonymous analysis and storage of the test data. The test result is immediately returned to the app displaying it to the user. Results: The limit of detection was 11.2 genome copies (95% CI) as determined by screening a tenfold dilution range of whole Leishmania donovani genomes using isothermal recombinase polymerase amplification (RPA). Pathogens considered for differential diagnosis were tested and no cross-reactivity was observed. For its diagnostic performance, DNA extracted from 170 VL and PKDL cases, comprising peripheral blood samples (VL, n = 96) and skin biopsies (PKDL, n = 74) from India (n = 108) and Bangladesh (n = 62), was screened. Clinical sensitivity and specificity were 88% and 91%, respectively. Conclusion: Minoo devices can offer a convenient, cheaper alternative to other molecular diagnostics. Its easy handling makes it ideal for use in low-resource settings to identify parasite burden. [ABSTRACT FROM AUTHOR]

Visceral leishmaniasis (VL), a fatal disease prevalent in more than 70 countries, poses significant health challenges, particularly in poor communities with limited access to healthcare. Vitamins and trace elements play a crucial role in immune function and may influence susceptibility to VL. This systematic review and meta-analysis aimed to assess the differences in serum vitamin and trace element levels in VL patients compared to healthy individuals. We conducted an extensive search of databases (PubMed, Embase, and Google Scholar) to identify potentially eligible articles published from inception to June 2023. Data extraction and quality assessment were carried out by two reviewers independently. RevMan software (version 5.4) was used for analysis. Standardized mean difference (SMD) with a confidence interval (CI) of 95% was used to summarize the findings. Ten studies comprising 546 VL patients and 535 controls were included in our study. The findings revealed significantly reduced serum retinol levels in VL patients in comparison to controls (SMD: − 0.67; 95% CI: [− 1.05, − 0.28]; p = 0.0008). Serum zinc levels were also substantially lower in VL patients, regardless of controls recruited from endemic (SMD: − 2.65; 95% CI: [− 3.86, − 1.44]; p < 0.0001) or non-endemic regions (SMD: − 1.99; 95% CI: [− 3.02, − 0.96]; p < 0.0002). However, VL patients exhibited significantly increased serum copper levels compared to controls (SMD: 2.51; 95% CI: [0.70, 4.32]; p = 0.007). Patients with VL had lower serum levels of zinc and retinol and higher levels of copper, indicating a possible role of these micronutrients in influencing VL susceptibility and progression. [ABSTRACT FROM AUTHOR]

Visceral leishmaniasis (VL) is an infectious disease caused by protozoa, Leishmania. During infection, the cell-mediated immunity of host gets impeded, so immunomodulation using natural products can be considered as an effective therapeutic strategy. The present study deals with the investigation of antileishmanial and immunostimulatory potential of hydro-ethanolic root extract of Rhodiola imbricata (HERERI) in inbred BALB/c mice. The secondary metabolites were analyzed quantitatively. To study the effect of HERERI on promastigotes, assays including inhibitory concentration (IC50) by trypan blue staining, cell cycle arrest by FACS were performed. The splenic parasite load was monitored after 14 days of treatment in BALB/c mice (500 and 1000 mg kg−1 b.wt.). The immunological responses after treatment were recorded in terms of DTH (delayed type of hypersensitivity) reactions, different T lymphocyte's subtypes and amounts of Th1/2 cytokines. Biochemical assays to analyze the function of liver and kidney were also done along with the histological studies. HERERI contains a considerable amount of flavonoids, phenols and terpenoids. The IC50 of 24.84 ± 0.52 and 46.65 ± 1.90 µg mL−1 against sensitive and resistant strain was obtained, respectively. It also arrested the cell cycle of promastigotes at sub G0-G1 phase. HERERI significantly decreased parasite load and up-scaled the DTH responses. It also polarized the Th1 dominating immune status by upregulation of IL-12, IFN-γ, and TNF-α cytokines and CD4+, CD8+ T cells. It caused no toxicity as biochemical tests and the histological sections were normal as compared to infected animals. Overall R. imbricata rescued mice from VL infection through immunostimulation. [ABSTRACT FROM AUTHOR]

Visceral leishmaniasis (VL) occurs due to the evolution, virulence, and adaptation of Leishmania, vector biology, host immune system evasion, and reservoir hosts. Parasitemia can be involved as a warning regarding the clinical severity of VL The present study aims to evaluate the relationship between parasitemia and the prognosis of individuals with VL. Blood and bone marrow samples from individuals with VL were analyzed to identify parasite and quantify or measure parasite burden. Individuals were classified in the clinical score model of risk of death by disease proposed by Coura-Vital et al. (PLoS Negl Trop Dis 8(12): e33742014, 2014). 39/74 individuals presented a better prognosis, and 35/74 individuals presented a worse prognosis. HIV + VL co-infection was present in 32 individuals, of which 12 were considered severe. The group aged 51 to 64 was classified as severe, with a decrease in leukocytes (p-value 0.0295) and neutrophils (p-value 0.0476). L. infantum DNA was identified in blood and bone marrow, in 69 individuals, and not detected in 5 individuals. The quantification of the parasite showed greater parasitemia in bone marrow (P = 0.0003) with an average of 4.70 × 104Leishmanias/mL about blood, with 0.29 × 104Leishmanias/mL. Individuals in the age group aged 51 to 64 co-infected with HIV + VL had higher parasitemia (p-value 0.0150) with 2.44 × 104Leishmanias/mL in blood and bone marrow than in the group aged 20 to 50. Parasitemia, measured by molecular biology in blood and bone marrow, was related to the worst clinical prognosis of VL in the age group aged 51 to 64. [ABSTRACT FROM AUTHOR]

Kinetoplastids are a group of flagellated protozoans including medically important parasites of the genus Trypanosoma and Leishmania. The corresponding diseases have afflicted humans for centuries. In an effort to combat kinetoplastid infections, a set of 21 chalcones was synthesized and evaluated for their in vitro anti-protozoal efficacy against Trypanosoma brucei, Trypanosoma brucei rhodesiense, Trypanosoma cruzi, and Leishmania infantum. To ensure safety, these compounds underwent a selectivity evaluation by assessing toxicity against a human lung fibroblast cell line. Compound K4 exhibited remarkable and selective trypanocidal activity against T. b. brucei with IC50 value of 0.31 ± 0.27 µM and T. b. rhodesiense with IC50 value of 0.96 ± 0.86 µM. Compound K9 also showed significant trypanocidal activity against T. b. brucei (IC50: 0.45 ± 0.14 µM) and T. b. rhodesiense (IC50: 0.93 ± 0.51 µM). In both compounds, electron withdrawing groups are appended to the styrenyl moiety. [ABSTRACT FROM AUTHOR]

A large proportion of HIV-coinfected visceral leishmaniasis (VL-HIV) patients exhibit chronic disease with frequent VL recurrence. However, knowledge on immunological determinants underlying the disease course is scarce. We longitudinally profiled the circulatory cellular immunity of an Ethiopian HIV cohort that included VL developers. We show that chronic VL-HIV patients exhibit high and persistent levels of TIGIT and PD-1 on CD8+/CD8- T cells, in addition to a lower frequency of IFN-γ+ TIGIT- CD8+/CD8- T cells, suggestive of impaired T cell functionality. At single T cell transcriptome and clonal resolution, the patients show CD4+ T cell anergy, characterised by a lack of T cell activation and lymphoproliferative response. These findings suggest that PD-1 and TIGIT play a pivotal role in VL-HIV chronicity, and may be further explored for patient risk stratification. Our findings provide a strong rationale for adjunctive immunotherapy for the treatment of chronic VL-HIV patients to break the recurrent disease cycle. A longitudinal study of the circulatory cellular immunity of individuals co-infected with recurrent visceral leishmaniasis and HIV suggests that T cell unresponsiveness underlies visceral leishmaniasis chronicity. [ABSTRACT FROM AUTHOR]

Visceral leishmaniasis-associated hemophagocytic lymphohistiocytosis (VL-HLH) is indistinguishable from those of HLH of other etiologies due to the overlap symptoms, posing a serious threat to life. In this study, we aimed to provide insights for early diagnosis and improve outcomes in pediatric patients with VL-HLH. We retrospectively analyzed the clinical and laboratory data of 10 pediatric patients with VL-HLH and 58 pediatric patients with Epstein–Barr virus-associated hemophagocytic lymphohistiocytosis (EBV-HLH). The median time from symptom onset to cytopenia in patients with VL-HLH and EBV-HLH was 11 days (interquartile range, 7–15 days) and five days (interquartile range, 3.75–9.25 days) (P = 0.005). Both groups showed liver injury and increased lactate dehydrogenase levels; however the levels of aspartate aminotransferase, alanine aminotransferase, direct bilirubin, and lactate dehydrogenase in patients with VL-HLH were significantly lower than those in patients with EBV-HLH (P < 0.05). The fibrinogen and triglyceride levels were almost normal in VL-HLH patients but were significantly altered in EBV-HLH cases (P < 0.05). The positive rate of first bone marrow microscopy examination, anti-rK39 IgG detection, and blood metagenomic next-generation sequencing was 50%, 100%, and 100%, respectively. After VL diagnosis, eight patients were treated with sodium stibogluconate and two were treated with liposomal amphotericin B. All the patients with VL-HLH recovered. Our study demonstrates that regular triglyceride and fibrinogen levels in pediatric patients with VL-HLH may help in differential diagnosis from EBV-HLH. VL-HLH is milder than EBV-HLH, with less severe liver injury and inflammatory responses, and timely treatment with antileishmanial agents is essential to improve the outcomes of pediatric patients with VL-HLH. [ABSTRACT FROM AUTHOR]

Visceral leishmaniasis (VL) is an infectious disease caused by Leishmania infantum. Clinically, VL evolves with systemic impairment, immunosuppression and hyperactivation with hypergammaglobulinemia. Although renal involvement has been recognized, a dearth of understanding about the underlying mechanisms driving acute kidney injury (AKI) in VL remains. We aimed to evaluate the involvement of immunoglobulins (Igs) and immune complexes (CIC) in the occurrence of AKI in VL patients. Fourteen VL patients were evaluated between early treatment and 12 months post-treatment (mpt). Anti-Leishmania Igs, CIC, cystatin C, C3a and C5a were assessed and correlated with AKI markers. Interestingly, high levels of CIC were observed in VL patients up to 6 mpt. Concomitantly, twelve patients met the criteria for AKI, while high levels of cystatin C were observed up to 6 mpt. Plasmatic cystatin C was positively correlated with CIC and Igs. Moreover, C5a was correlated with cystatin C, CIC and Igs. We did not identify any correlation between amphotericin B use and kidney function markers in VL patients, although this association needs to be further explored in subsequent studies. Our data reinforce the presence of an important renal function impairment during VL, suggesting the involvement of Igs, CIC, and C5a in this clinical condition. [ABSTRACT FROM AUTHOR]

Purpose: Visceral leishmaniasis (VL) is a systemic and parasitic disease that is usually fatal if left untreated. VL is endemic in different parts of Iran and is caused mainly by Leishmania infantum. This study aimed to recognition immunoreactive proteins in amastigote-like and promastigote stages of L. infantum (Iranian strain) by antibodies present in the sera of VL patients. Methods: Total protein extract from amastigote-like and promastigote cells was separated by two-dimensional electrophoresis (2DE). To detect the immunoreactive proteins, 2DE immunoblotting method was performed using different pools of VL patients' sera. Results: Approximately 390 and 430 protein spots could be separated in 2DE profiles of L. infantum amastigote-like and promastigote stages, respectively. In immunoblotting method, approximately 295 and 135 immunoreactive proteins of amastigotes-like reacted with high antibody titer serum pool and low antibody titer serum pool, respectively. Approximately 120 and 85 immunoreactive proteins of promastigote extract were recognized using the high antibody titer sera pool and low antibody titer sera, respectively. Conclusion: The present study has recognized a number of antigenic diversity proteins based on the molecular weight and pH in amastigote-like and promastigote stages of L. infantum. These results provide us a new concept for further analysis development in the field of diagnosis biomarkers and vaccine targets. [ABSTRACT FROM AUTHOR]

Visceral leishmaniasis (VL) requires diagnostic assays to complement clinical suspicion. However, there is no standardization of a diagnostic flow using available assays. This study aimed to evaluate the performance of parasitological, molecular, and serological assays for diagnosing VL and propose a diagnostic flow based on performance, practicality, and invasiveness. We conducted a study of 10-year (2010–2020) routine diagnoses of VL at the Brazilian National Reference Laboratory. We propose a diagnostic flow where individuals suspected of VL are initially screened for antibodies using an immunochromatographic test (ICT) with rK39 antigen on the nitrocellulose membrane. This is followed by a blood polymerase chain reaction (PCR) for Leishmania sp. kDNA and direct parasitological exam and/or PCR in bone marrow aspirate. A positive result in any of these assays can define a VL case. If clinical suspicion persists in negative individuals, the diagnostic flow should be repeated. The proposed flow has the potential to standardize and improve the diagnosis of VL. It reduces the need for invasive tests without compromising diagnostic accuracy. [ABSTRACT FROM AUTHOR]

Since 1999, the number of asymptomatic leishmaniasis cases has increased continuously in Thailand, particularly among patients with HIV who are prone to develop symptoms of cutaneous and visceral leishmaniasis further. The asymptomatic infection could play a key role in Leishmania transmission and distribution. Understanding population structure and phylogeographic patterns could be crucially needed to develop effective diagnoses and appropriate guidelines for therapy. In this study, genetic variation and geographic distribution of the Leishmania/HIV co-infected population were investigated in endemic northern and southern Thailand. Interestingly, Leishmania orientalis was common and predominant in these two regions with common regional haplotype distribution but not for the others. Recent population expansion was estimated, probably due to the movement and migration of asymptomatic individuals; therefore, the transmission and prevalence of Leishmania infection could be underestimated. These findings of imbalanced population structure and phylogeographic distribution patterns provide valuable, insightful population structure and geographic distribution of Leishmania/HIV co-infection to empower prevention and control of transmission and expansion of asymptomatic leishmaniasis. [ABSTRACT FROM AUTHOR]

Tegumentary leishmaniasis (TL) is the main clinical manifestation of leishmaniasis, and it can cause the infected hosts to self-healing cutaneous lesions until mutilating scars in mucosal membranes, particularly in the nose and throat. The treatment against disease presents problems, and the diagnosis is hampered by variable sensitivity and/or specificity of the tests. In this context, the development of prophylactic vaccines could be considered as a strategy to control the disease. Previously, we showed that the recombinant LiHyp1 protein plus adjuvant protected mice from infection with Leishmania infantum, which causes visceral leishmaniasis. In the present study, we tested whether rLiHyp1 could induce protection against infection with L. amazonensis, a parasite species able to cause TL. We immunized BALB/c mice with rLiHyp1 plus saponin (rLiHyp1/S) or incorporated in micelles (rLiHyp1/M) as adjuvants and performed parasitological and immunological evaluations before and after infection. Results showed that after in vitro stimulation from spleen cell cultures using rLiHyp1 or a Leishmania antigenic extract (SLA), rLiHyp1/S and rLiHyp1/M groups developed a Th1-type immune response, which was characterized by high levels of IFN-γ, IL-2, TNF-α and IL-12 cytokines, nitrite, and IgG2a isotype antibodies when compared to values found in the control (saline, saponin, micelles alone) groups, which showed higher levels of anti-SLA IL-4, IL-10, and IgG1 antibodies before and after challenge. In addition, mice receiving rLiHyp1/S or rLiHyp1/M presented significant reductions in the lesion average diameter and parasite load in the infected tissue and internal organs. Blood samples were collected from healthy subjects and TL patients to obtain PBMC cultures, which were in vitro stimulated with rLiHyp1 or SLA, and results showed higher lymphoproliferation and IFN-γ production after stimulus using rLiHyp1, as compared to values found using SLA. These results suggest that rLiHyp1 plus adjuvant was protective against experimental TL and could also be considered for future studies as a vaccine candidate against human disease. [ABSTRACT FROM AUTHOR]

Canine visceral leishmaniasis (CVL) is a significant vector-borne Meta zoonotic disease caused by agents of the L. donovani complex. The disease is transmitted by the bite of phlebotomine female sandflies of the genera Phlebotomus and Lutzomyia in the old and new worlds, respectively. This study was conducted to assess the knowledge, attitude, and practices of the residents about CVL based on the One Health concept in two towns of the Oromia Region, Ethiopia. A community-based cross-sectional study was conducted between October 2019 and September 2020, using an interview questionnaire as the study instrument. The study participants were selected through a simple random sampling method. Pearson's Chi-square and logistic regression tests were used to evaluate the association between the study participants' knowledge, attitude, and practices toward CVL and possible risk factors. The study included a total of 400 participants, and the results indicated that 77.25% had good knowledge, 60.5% had a favorable attitude, and 59.25% had good practices toward CVL. The town of Ejaji and dog ownership were significantly associated with good knowledge (p = 0.001), and attitude (p = 0.001) towards CVL, while having a dog (p = 0.001), having a diploma (p = 0.019) or degree and above (0.018), being divorced or widowed (0.012), and being Oromo (p = 0.013) were all significantly associated with good CVL practice. Most participants in both study areas had good knowledge but moderate attitudes and practices toward CVL. Therefore, it is crucial to undertake comprehensive community health education and awareness programs of zoonotic visceral leishmaniasis and its vectors based on the One Health concept through various means. [ABSTRACT FROM AUTHOR]

The leishmanin skin test was used for almost a century to detect exposure and immunity to Leishmania, the causative agent of leishmaniasis, a major neglected tropical disease. Due to a lack of antigen used for the intradermal injection, the leishmanin skin test is no longer available. As leishmaniasis control programs are advancing and new vaccines are entering clinical trials, it is essential to re-introduce the leishmanin skin test. Here we establish a Leishmania donovani strain and describe the production, under Good Laboratory Practice conditions, of leishmanin soluble antigen used to induce the leishmanin skin test in animal models of infection and vaccination. Using a mouse model of cutaneous leishmaniasis and a hamster model of visceral leishmaniasis, soluble antigen induces a leishmanin skin test response following infection and vaccination with live attenuated Leishmania major (LmCen-/-). Both the CD4+ and CD8+ T-cells are necessary for the leishmanin skin test response. This study demonstrates the feasibility of large-scale production of leishmanin antigen addressing a major bottleneck for performing the leishmanin skin test in future surveillance and vaccine clinical trials. As leishmaniasis control programs and new vaccines are advancing, it is necessary to re-introduce the leishmanin skin test to monitor transmission and immunity. This study describes the generation and validation of a new leishmanin skin test antigen for future re-introduction into endemic countries. [ABSTRACT FROM AUTHOR]

Feline leishmanial infection is reported worldwide, but the epidemiological role of domestic cats in the leishmaniasis cycle remains unclear, and cats might act as cryptic reservoir hosts in endemic areas with no feline leishmaniosis cases. Considering that, a serological screening for anti-Leishmania spp. antibodies was performed by indirect immunofluorescence antibody test (IFAT) in 389 necropsied cats' serum samples from a new visceral leishmaniasis transmission area with no feline leishmanial infection reported to unveil if the cats are being exposed to the parasite. The overall seroprevalence for Leishmania spp. was 11.05% (43/389). No association was found between sex, neutering status, age group, breed, coat length, feline immunodeficiency virus (FIV) infection, and Leishmania spp. antibody detection. A positive association was found with coat color (cats within the orange spectrum with white [particolor]) (OR = 2.47, CI 95% 1 – 6.13, P = 0.044) and a negative association (OR = 0.38, CI 95% 0.18 – 0.79, P = 0.01) between feline leukemia virus (FeLV) infection and IFAT positivity for Leishmania spp. Therefore, it is concluded that the seroprevalence found was greater than 10%, indicating contact of the protozoan with cats in the region served. [ABSTRACT FROM AUTHOR]

We aimed to estimate the diagnostic latency of patients with visceral leishmaniasis (VL). A monocentric retrospective observational study was conducted including all confirmed cases of VL diagnosed from January 2005 to March 2022. Epidemiological and clinical characteristics of patients with VL were collected. The diagnostic latency was defined as the number of days between the first contact with a health-care provider for signs and/or symptoms referable to VL and the laboratory diagnosis of leishmaniasis. Twenty-four cases of VL were included in the study, mostly male (75%) and Italians (79.2%), with a median age of 40 years [Inter Quartile Range (IQR 30–48)]. Fourteen (58.3%) VL cases were people living with HIV (PLWH) and 4 (16.6%) subjects were on immunosuppressive therapy. For VL the median diagnostic latency was 54 days (IQR 28–162). The shorter diagnostic latency was observed in PLWH [31 days (IQR 20–47)] followed by immunocompetent patients [160 days (IQR 133–247)] and those on immunosuppressive therapy [329 days (IQR 200–678)]. Twelve patients (50%) reported at least one medical encounter before the diagnosis of VL and 6 patients received a wrong therapy. Diagnostic delay in VL was significant in patients under immune suppressive treatment. [ABSTRACT FROM AUTHOR]

Leishmaniasis is a commonly neglected disease present in tropical and subtropical countries, affecting 1 billion people. Visceral Leishmaniasis (VL) is the most severe form and can lead to death if left untreated. In this work, we apply deep learning techniques to detect VL in humans through images of slides from the parasitological examination (microscopy) of the bone marrow, aiding in an automatic and accurate diagnosis. This work investigates five deep learning architectures combined with preprocessing, data augmentation, and fine-tuning techniques to detect this disease in images. We compared our results with five related state-of-the-art works, which showed that the proposed classification method surpassed them in all metrics. We achieve an Accuracy of 98.7%, an F1-Score of 98.7%, and a Kappa of 98.7%. Therefore, we demonstrated that trained deep learning models with microscopic slide imaging of bone marrow biological material could precisely help the specialist detect VL in humans. [ABSTRACT FROM AUTHOR]

In this serum panel–based study, we evaluated the accuracy of serological tests originally developed for visceral leishmaniasis (VL), for diagnosis of mucosal leishmaniasis (ML). A total of five tests were evaluated, four of which are registered at the National Agency of Sanitary Surveillance (Agência Nacional de Vigilância Sanitária-ANVISA) (RIDASCREEN® Leishmania Ab from R-Biopharm AG., Leishmania ELISA IgG + IgM from Vircell S.L., IFI Leishmaniose Humana—BioManguinhos, and IT-LEISH® from Bio-Rad Laboratories, Inc.), and the other a direct agglutination test (DAT-LPC) prototype kit developed at Fiocruz. The panel was composed of 40 serum samples from patients with confirmed ML and 20 from patients with mucosal involvement and negative parasitological/molecular tests for leishmaniasis and confirmation of another etiology. All cases were treated from 2009 to 2016 in a referral center for leishmaniasis in Belo Horizonte, Minas Gerais, Brazil (Instituto René Rachou, Fiocruz). Diagnostic accuracy, based on the cut-off point for VL diagnosis, was 86.2% with RIDASCREEN® Leishmania Ab, 73.3% with Leishmania ELISA IgG + IgM, and 66.7% with IFI Leishmaniose Humana, while IT-LEISH® and DAT-LPC had the lowest accuracy (38.3%), despite high specificity (100% and 95%, respectively). New cut-off points defined with sera from ML patients improved accuracy from 86.2 to 89% (p = 0.64) and 73.3 to 88% (p = 0.04) for RIDASCREEN® Leishmania Ab and Leishmania ELISA IgG + IgM, respectively. Moreover, these tests presented greater sensitivity and immunoreactivity in patients with moderate/severe clinical ML forms. The data of this study suggest that ELISA assays can contribute to laboratory diagnosis, especially for patients with moderate or severe mucosal involvement. [ABSTRACT FROM AUTHOR]

Purpose: In Turkey, the main causative agent of visceral leishmaniasis (VL) is Leishmania. infantum and the main causative agent of cutaneous leishmaniasis (CL) is Leishmania tropica. In this study, we aimed to discuss the possible mechanisms, clinical aspects, and threat of visceralizing L. tropica. Methods: This study includes seven cases of VL caused by L. tropica.Five patients were male (71%) and four were adults (57%). Results: All the VL patients complained of fever and splenomegaly. Fatigue, pancytopenia, and hepatomegaly were present in six patients each (86%), while weight loss and gastrointestinal system (GIS) symptoms were present in 5 patients (71%). Conclusions: In this study, we have evaluated seven cases of visceralized L. tropica (VLT) in the context of the changing leishmaniasis epidemiology in Turkey. We have evaluated the possible mechanisms of visceralization; inter- and intraspecies genetic exchange with all the old world leishmaniasis agents present in the region, stress induced by inappropriate use of drugs, and possible ongoing adaptation mechanisms of Leishmania spp. The threat posed by VLT is significant as L. tropica is the most widespread and most common cause of leishmaniasis in Turkey. We do not know the vectorial capacity of the sand flies for the transmission of VLT strains or if these strains are in circulation in Turkey. Future studies should be carried out to investigate these issues as the transition of L. tropica from a mild disease-causing agent to a mortal one poses a significant public health concern for Turkey and Europe. [ABSTRACT FROM AUTHOR]

Objective: Phlebotomine sand flies are the primary vectors of leishmaniasis; visceral form of disease is transmitted mainly by species belonging to Larroussius and Adlerius subgenus. Species identification of some females in Larroussius subgenus is not easy due to the high similarity. Accurate species identification enables control operations to target only main vectors and increase understanding of ecological needs, bionomic characteristics, and behavioral traits. The purpose of current study was to use two approaches based on internal and external morphological characters to identify the wild caught of female specimens belonged to Larroussius subgenus and investigate Leishmania infection. Methods: Totally, 128 specimens belonging to Larroussius' subgenus were collected from a VL focus in northwestern Iran, species differentiation was done by two approaches proposed in literature: (1) characters of pharyngeal armature, number of Spermathecal segment, length of spermathecal neck, palpal and ascoid formulae; (2) blindly on the basis of the shape of the base of spermathecal duct. Their potential infection to Leishmania was investigated by kDNA-Nested-PCR. Results: The results of species identification were consistent based on two methods. Among three identified species, Phlebotomus perfiliewi was found to be the dominant species, followed by Ph. neglectus and Ph. tobbi. Two specimens of Ph. perfiliewi were found to be infected by Leishmania infantum, which emphasizes the role of this species in VL transmission in study area. Conclusion: It is suggested that combination of the characters used here be considered for species identification of female of Larroussius subgenus to take advantage of maximum characters, especially where species present sympatrically. [ABSTRACT FROM AUTHOR]

Leishmania donovani is the causative organism for visceral leishmaniasis. Although this parasite was discovered over a century ago, nothing is known about role of potassium channels in L. donovani. Potassium channels are known for their crucial roles in cellular functions in other organisms. Recently the presence of a calcium-activated potassium channel in L. donovani was reported which prompted us to look for other proteins which could be potassium channels and to investigate their possible physiological roles. Twenty sequences were identified in L. donovani genome and subjected to estimation of physio-chemical properties, motif analysis, localization prediction and transmembrane domain analysis. Structural predictions were also done. The channels were majorly α-helical and predominantly localized in cell membrane and lysosomes. The signature selectivity filter of potassium channel was present in all the sequences. In addition to the conventional potassium channel activity, they were associated with gene ontology terms for mitotic cell cycle, cell death, modulation by virus of host process, cell motility etc. The entire study indicates the presence of potassium channel families in L. donovani which may have involvement in several cellular pathways. Further investigations on these putative potassium channels are needed to elucidate their roles in Leishmania. [ABSTRACT FROM AUTHOR]

Leishmaniasis is a protozoal infection transmitted by sandfly vector; there are three main types of leishmaniasis: cutaneous leishmaniasis (CL), mucocutaneous leishmaniasis (MCL), and visceral leishmaniasis (VL). Herein, we present a case of endonasal mucocutaneous leishmaniasis in a 34-year-old Yemeni patient who presented with disfiguring nasal swelling associated with fever, a swab from the lesion and direct microscopic examination proved to have mucocutaneous leishmaniasis; he was successfully treated with intravenous antimonial stibogluconate, 1 month after treatment biopsy from the lesion revealed cytotoxic T-cell lymphoma, the purpose is to alert the physicians and otolaryngologists to consider leishmaniasis in the differential diagnosis of nasal granulomas and also to highlight the importance of early diagnosis of cancer in survivors of cutaneous leishmaniasis, especially in areas where cutaneous leishmaniasis is still highly prevalent, as the chronic local inflammation may disfigure the face if not recognized early and adequately treated; also, early cancer diagnosis can prevent mortality [ABSTRACT FROM AUTHOR]

Context: Leishmaniasis is a group of vector-borne infectious diseases caused by over 20 pathogenic Leishmania species that are endemic in many tropical and subtropical countries. The emergence of drug-resistant strains, the adverse side effects of anti-Leishmania drugs, and the absence of a preventative vaccination strategy threaten the sensitive population. Recently, many groups of researchers have exploited the field of reverse vaccinology to develop vaccines, focusing chiefly on inducing immunity against either visceral or cutaneous leishmaniasis. Methods: This present work involves retrieving twelve experimentally validated leishmanial antigenic protein sequences from the UniProt database, followed by their antigenicity profiling employing ANTIGENpro and Vaxijen 2.0 servers. MHC-binding epitopes for the same were predicted using both NetCTL 1.2 and SYFPEITHI servers, while epitopes for B cell were computed using ABCpred and BepiPred 2.0 servers. The screened epitopes with significantly higher scores were utilized for designing the vaccine construct with appropriate linkers and natural adjuvant. The secondary and tertiary structures of the synthetic peptide were determined by conditional random fields, shallow neural networks, and profile-profile threading alignment with iterative structure assembly simulations, respectively. The 3-D vaccine model was validated through CASP10-tested refinement and the MolProbity web server. Molecular docking and multi-scale normal mode analysis simulation were performed to analyze the best vaccine-TLR complex. Finally, computational immune simulation findings revealed promising cellular and humoral immune responses, suggesting that the engineered chimeric peptide is a potential broad-spectrum vaccine against visceral and cutaneous leishmaniasis. [ABSTRACT FROM AUTHOR]

Visceral leishmaniasis is an infectious disease caused by the protozoan Leishmania. Due to the lack of vaccines and the limitations of current treatment (toxicity, serious adverse effects, and high cost), new therapeutic options must be developed. This work aims to perform a systematic review focusing on studies that use nanoparticles to treat visceral leishmaniasis, with in vivo tests to evaluate the antileishmanial effect. The search was carried out between January 2011 and August 2021 in the PubMed and Web of Science databases, using the keywords "visceral leishmaniasis and nanoparticles and in vivo." The articles were selected through inclusion and exclusion criteria. After the analysis, 26 articles were selected containing polymeric, lipidic, metallic, and inorganic nanoparticles. As for the nanometric size, the polymeric and lipid nanoparticles showed size ranges from 138 to 377.9 nm and 103.7 to 266.3 nm, respectively. The inorganic nanoparticles had a size of around 140 nm. Also, the metallic nanoparticles showed sizes of 58.6, 134.3, and 142.7 nm. In addition, amphotericin B was the most used drug. In some studies, drug co-encapsulation was performed, which is a future trend, useful to increase the therapeutic efficacy of the treatment. The studies showed that the use of nanoparticles for drug delivery was able to reduce the parasite burden and toxicity. Nanoparticles can increase the efficacy and safety of the treatment, suggesting a promising option for visceral leishmaniasis treatment. [ABSTRACT FROM AUTHOR]

Human visceral leishmaniasis (HVL) is a neglected disease that occurs in 98 countries on five continents, and it is endemic in tropical and subtropical regions. In South America, the etiological agent of HVL is Leishmania infantum (syn. Leishmania chagasi), mainly transmitted through the bite of an infected sandfly female from the genus Lutzomyia. In American HVL endemic areas, the occurrence of asymptomatic infection is common, which contributes to the possibility of L. infantum transmission during a blood transfusion. To know the prevalence of L. infantum asymptomatic infection in blood donors from the microregion of Adamantina, we investigated 324 peripheral blood samples from donors through immunofluorescence (IFAT) and PCR–RFLP techniques. Seven blood samples (2.16%) tested positive for Leishmania by IFAT, and from those, six presented positive results by PCR (85.71%), which were later identified as L. infantum by RFLP. The presence of L. infantum in the peripheral blood of blood donors supported the hypothesis of transmission by blood transfusion and points to the need to include tests for visceral leishmaniasis in blood bank screening tests and pre-storage measures, especially in endemic areas to prevent the exponential increase of HVL by blood transfusion. [ABSTRACT FROM AUTHOR]

Parasite of genus Leishmania viz. L. donovani and L. infantum cause visceral leishmaniasis (VL) or Kala-azar, systemic disease with significant enlargement of the liver and spleen, weight loss, anemia, fever and immunosuppression. The silent expansion of vectors, reservoir hosts and resistant strains is also of great concern in VL control. Considering all these issues, the present study focused on in vitro and in vivo antileishmanial screening of ellagic acid (EA) against L. donovani. The in vitro study was performed against the protozoan parasite L. donovani and a 50% inhibitory concentration was calculated. The DNA arrest in the sub-G0/G1 phase of the cell cycle was studied. In vivo studies included the assessment of parasite burden and immunomodulation in response to treatment of ellagic acid in BALB/c mice. The levels of Th1 and Th2 cytokines and isotype antibodies were assessed in different groups of mice. EA showed in vitro parasiticidal activity with IC50 18.55 µg/mL and thwarted cell-cycle progression at the sub-G0/G1 phase. Administration of ellagic acid to the BALB/c mice reported diminution of splenic and hepatic parasite burden coupled with an expansion of CD4+ and CD8+ T lymphocytes. EA further potentiated a protective immune response with augmentation of Th1 type immune response evidenced by elevation of serum IgG2a levels and DTH response. EA was reported to be safe and non-toxic to the THP-1 cell line as well as to the liver and kidneys of mice. These findings endorse the therapeutic potential of EA with significant immunomodulation and can serve as a promising agent against this debilitating parasitic disease. [ABSTRACT FROM AUTHOR]

Visceral leishmaniasis is a major, life-threatening parasitic disease that still remains a serious public health problem in Ethiopia. Understanding the epidemiological, clinical, and hematological profiles of visceral leishmaniasis patients is important for implementing evidence-based control strategies. It is also important for early treatment and to decrease the mortality rate from the disease. Therefore, this study was aimed at assessing the epidemiological, clinical, and hematological profiles of visceral leishmaniasis among patients visiting Tefera Hailu Memorial Hospital, Northeast Ethiopia. A retrospective study was conducted at Tefera Hailu Memorial Hospital from September 2017 to August 2021. Data were collected from the medical records of suspected patients who were tested by the rK39 rapid diagnostic by strictly following standard operating procedures. The data was summarized using Microsoft Excel and analyzed using SPSS 26 version software. Descriptive statistics were used to describe the epidemiological, clinical, and hematological profiles of visceral leishmaniasis patients. A p-value < 0.05 was considered statistically significant. The overall positivity rate for visceral leishmaniasis was 23.4% (132/564). The result of this study indicated a fluctuating yet declining trend in VL over the past 4 years. From a total of 132 VL confirmed cases, the numbers of cases were highest among males (78.0%), those 15–29 years of age (37.1%), and urban residents (89.4%). Furthermore, Abergele (11.0%), Sehala (6.0%), and Ziquala (5.0%) districts had the highest number of VL cases. The major clinical presentations of patients were fever (96.2%), splenomegaly (94.7%), and general weakness (80.3%). With regard to hematological profiles, the most common findings were anemia (86.4%), thrombocytopenia (81.8%), leucopenia (78.8%), neutropenia (74.2%), and pancytopenia (71.2%). In the study area, the VL positivity rate was high. Our findings also concluded that VL causes significant alterations in clinical and hematological parameters. Therefore, the zone health office and other concerned stakeholders should strengthen evidence-based control programs for VL. [ABSTRACT FROM AUTHOR]

Leishmaniasis is a parasitic infection expressing different clinical phenotypes. Visceral leishmaniasis (VL) is considered an opportunistic infection among people with human immunodeficiency virus (HIV). The objective of this review was to identify published data on the prevalence of Leishmania spp. infection among PWH and to define particular determinants that affect critically the epidemiological characteristics of VL-HIV coinfection and, potentially, its burden on public health. Two independent reviewers conducted a systematic literature search until June 30, 2022. Meta-analyses were conducted using random-effects models to calculate the summary prevalence and respective 95% confidence intervals (CI) of leishmaniasis among PWH. Meta-regression analysis was performed to investigate the impact of putative effect modifiers, such as the mean CD4 cell count, on the major findings. Thirty-four studies were eligible, yielding a summary prevalence of 6% (95%CI, 4–11%) for leishmaniasis (n = 1583) among PWH (n = 85,076). Higher prevalence rates were noted in Asia (17%, 95%CI, 9–30%) and America (9%, 95%CI, 5–17%) than in Europe (4%, 95%CI, 2–8%). Prevalence rates were significantly mediated by the age, sex, and CD4 cell count of participants. Heterogeneity remained significant in all meta-analyses (p < 0.0001). In the majority of included studies, people were coinfected with HIV and Leishmania species associated with VL, as opposed to those associated with cutaneous leishmaniasis. No sign of publication bias was shown (p = 0.06). Our summary of published studies on leishmaniasis among PWH is important to provide prevalence estimates and define potential underlying factors that could guide researchers to generate and further explore specific etiologic hypotheses. [ABSTRACT FROM AUTHOR]

The utility of andrographolide (AN) in visceral leishmaniasis (VL) and cutaneous leishmaniasis (CL) is limited owing to poor solubility, hindered permeation, and unstable structure under physiological conditions. The present study mainly focuses on synthesizing of andrographolide-Soya-L-α-phosphatidyl choline (ANSPC) complex in ethanol and its characterization using various spectral and analytical techniques. Results from FT-IR, 1H NMR, ROSEY, and in silico docking techniques suggest ANSPC complex formation due to inter-molecular interaction between the hydrophilic head of SPC and hydroxyl group of AN present at 24th position. ANSPC complex demonstrated the solubility of 113.93 ± 6.66 μg/mL significantly (P < 0.05) greater than 6.39 ± 0.47 μg/mL of AN. The particle size of ANSPC complex was found to be 182.2 ± 2.69 nm. The IC50 value of AN suspension (PBS, pH ~ 7.4) at 24, 48, and 72 h against Leishmania donovani (L. donovani) was noticed to be 32.76 ± 4.53, 20.87 ± 2.37, and 17.71 ± 3.06 μM/mL, respectively. Moreover, augmented aqueous solubility of ANSPC complex led to significant (P < 0.05) reduction in IC50 value, i.e., 25.02 ± 4.35, 11.31 ± 0.60, and 8.33 ± 2.71 μM/mL at 24, 48, and 72 h, respectively. The IC50 values for miltefosine were noted to be 9.84 ± 2.65, 12.13 ± 7.26, and 6.56 ± 0.61 μM/mL at similar time periods. Moreover, ANSPC complex demonstrated augmented cellular uptake at 24 h as compared to 6 h in L.donovani. We suppose that submicron size and phospholipid-mediated complexation might have endorsed the permeation of ANSPC complex across the plasma membrane of L.donovani parasite by transport mechanisms such as P-type ATPase. ANSPC complex warrants further in-depth in vivo studies under a set of stringent parameters for translating the product into a clinically viable form. [ABSTRACT FROM AUTHOR]

Leishmaniasis is one of the top neglected tropical diseases with significant morbidity and mortality in low and middle-income countries (LMIC). However, this disease is also spreading in the developed world. Currently, there is a lack of effective strategies to control this disease. Vaccination can be an effective measure to control leishmaniasis and has the potential to achieve disease elimination. Recently, we have generated centrin gene-deleted new world L. mexicana (LmexCen−/−) parasites using CRISPR/Cas9 and showed that they protect mice against a homologous L. mexicana infection that causes cutaneous disease. In this study, we tested whether LmexCen−/− parasites can also protect against visceral leishmaniasis caused by L. donovani in a hamster model. We showed that immunization with LmexCen−/− parasites is safe and does not cause lesions. Furthermore, such immunization conferred protection against visceral leishmaniasis caused by a needle-initiated L. donovani challenge, as indicated by a significant reduction in the parasite burdens in the spleen and liver as well as reduced mortality. Similar control of parasite burden was also observed against a sand fly mediated L. donovani challenge. Importantly, immunization with LmexCen−/− down-regulated the disease promoting cytokines IL-10 and IL-4 and increased pro-inflammatory cytokine IFN-γ resulting in higher IFN-γ/IL-10 and IFN-γ/IL4 ratios compared to non-immunized animals. LmexCen−/− immunization also resulted in long-lasting protection against L. donovani infection. Taken together, our study demonstrates that immunization with LmexCen−/− parasites is safe and efficacious against the Old World visceral leishmaniasis. [ABSTRACT FROM AUTHOR]

Leishmania donovani and Leishmania infantum are closely related species. However, the former is considered the causative agent for anthroponotic visceral leishmaniasis (AVL), while the latter is known to be responsible for zoonotic visceral leishmaniasis (ZVL) with dogs as the main reservoir host. Although molecular detection of L. donovani from naturally infected dogs has been reported in AVL endemic areas, the experimental infection of dogs with this species is very limited. Here, we constructed an experimental canine visceral leishmaniasis (CVL) model with L. donovani infection using beagle dogs. During an observation period of 8 months after parasite inoculation, few clinical symptoms were observed in the three inoculated dogs. The overall hematological and biochemical data of the dogs showed normal levels, and there were no remarkable changes in the peripheral CD4+, CD8+, CD25+, or FoxP3+ T cell populations. Liver biopsy sampling was conducted to monitor the parasite burden in the liver. A similar pattern of the amount of mitochondrial kinetoplast DNA was observed in the peripheral blood and liver by real-time PCR analysis. In addition, parasite antigens were detected from the liver biopsy sections by immunohistochemical analysis, further supporting the existence of parasites in the liver. These results showed a subclinical CVL model for L. donovani in beagle dogs with a similar kinetics of parasite burden in the peripheral blood and liver. [ABSTRACT FROM AUTHOR]

Purpose: Leishmaniasis is a neglected infectious disease affecting millions of people worldwide. Visceral leishmaniasis (VL), caused by Leishmania infantum and Leishmania donovani, is one of the main clinical forms of the disease and fatal if not treated promptly and properly. Despite being available for the last 70 years, current drugs used in the treatment of leishmaniasis have serious problems as they have high toxicity, require long-term administration and cause serious side-effects, leading to the emergence of resistant and relapse cases. Therefore, there is an urgent need for the discovery of novel antileishmanial molecules and the development of new treatment regimens. The drug used for chemotherapy of B-cell malignancies, Ibrutinib, an inhibitor of Bruton's Tyrosine Kinase (BTK), can offer a new therapeutic perspective due to the functions of BTK on intracellular signaling mechanism of macrophages, which are the primary resident cell for Leishmania. Hence, the study aimed to evaluate ibrutinib as a potential anti-Leishmanial drug. Method: In this study, we evaluated the antileishmanial effect of Ibrutinib by in vitroL. infantum infection model using macrophages, with cell viability assay, parasite rescue assay, real-time qPCR. Results: We showed that Ibrutinib was significantly more effective than the Glucantime against L. infantum. In addition, our data revealed that Ibrutinib inhibited parasite growth and load without impairing macrophage viability. Conclusions: Consequently, due to its efficacy and safety, Ibrutinib may be a promising candidate for the treatment of VL caused by L. infantum as a host-targeted drug. [ABSTRACT FROM AUTHOR]

In the absence of adequate diagnosis and treatment, leishmaniasis remains a major public health concern on a global scale. Drug resistance remains a key obstacle in controlling and eliminating visceral leishmaniasis. The therapeutic gap due to lack of target-specific medicine and vaccine can be minimized by obtaining parasite's genomic information. This study compared whole-genome sequence of paromomycin-resistant parasite (K133PMM) developed through in vitro adaptation and selection with sensitive Leishmania clinical isolate (K133WT). We found a large number of upstream and intergenic gene variations in K133PMM. There were 259 single nucleotide polymorphisms (SNPs), 187 insertion-deletion (InDels), and 546 copy number variations (CNVs) identified. Most of the genomic variations were found in the gene's upstream and non-coding regions. Ploidy estimation revealed chromosome 5 in tetrasomy and 6, 9, and 12 in trisomy, uniquely in K133PMM. These contain the genes for protein degradation, parasite motility, autophagy, cell cycle maintenance, and drug efflux membrane transporters. Furthermore, we also observed reduction in ploidy of chromosomes 15, 20, and 23, in the resistant parasite containing mostly the genes for hypothetical proteins and membrane transporters. We chronicled correlated genomic conversion and aneuploidy in parasites and hypothesize that this led to rapid evolutionary changes in response to drug induced pressure, which causes them to become resistant. [ABSTRACT FROM AUTHOR]

Sand flies are hematophagous dipterans and are considered vectors of the parasites belonging to the genus Leishmania, which can infect mammals, including humans, and subsequently cause leishmaniasis. Rio Grande do Sul State (RS), southern Brazil, was considered free of this disease for the last few decades. However, several autochthonous cases of visceral leishmaniasis (VL) have been recorded in different municipalities in recent years. Since 2017, cases of canine visceral leishmaniasis (CVL) have been reported in the municipality of Santa Maria, located in the central region of RS. In 2021, two cases of human VL were confirmed in the city, resulting in one death. To capture the possible vectors involved in the disease transmission cycle, Centers for Disease Control (CDC) modified light traps were installed in the peridomestic residential areas. These points were situated in two city districts where cases of CVL had been reported from January 2021 to January 2022. In the 41 sample collections of sand flies, 9 specimens of Lutzomyia longipalpis were identified, confirming the municipality as an area of VL transmission, according to Brazilian sanitary standards. The spread of Leishmania spp. has been occurring efficiently, resulting in a negative impact on the One Health. This study reports the first detection of L. longipalpis in the central region of RS. [ABSTRACT FROM AUTHOR]

In Brazil, visceral leishmaniasis (VL) has been expanding and urbanizing, mainly in non-endemic areas such as the State of Rio Grande do Sul. Considering that infected dogs are the main reservoirs of VL in urban areas, the present study aimed to evaluate the propagation of canine visceral leishmaniasis (CVL) infection from an unaffected region in transition to a VL transmission area. For this, 1159 and 1087 samples of canine serum from 2015 and 2021, respectively, were analyzed, using the indirect immunofluorescence test. In addition, necropsy reports between 2007 and 2021 were evaluated. The results showed a prevalence of anti-Leishmania spp. antibodies of 7.5% in the samples from 2015, while in 2021 samples, it was 23.5%, with an incidence of 0.4 cases per 100 dogs. It is noteworthy that in 2007, there was no record of CVL as the cause of death in the pathological reports, and in 2021, 41 diagnoses were made with the protozoan being a determinant of the death of the animal. These values indicate an increasing trend in the prevalence and incidence coefficients of CVL. The results of this study allowed us to verify the spread of the disease from an unaffected region to a transmission area of the agent, as well as provide subsidies for health authorities to implement improvements in the CVL control program in the municipality, to mitigate the emergence of human cases of the disease. [ABSTRACT FROM AUTHOR]

To date, sand flies (Phlebotominae) are the only recognized biological vectors of Leishmania infantum, the causative agent of human visceral leishmaniasis, which is endemic in the Mediterranean basin and also widespread in Central and South America, the Middle East, and Central Asia. Dogs are the main domestic reservoir of zoonotic visceral leishmaniasis, and the role of secondary vectors such as ticks and fleas and particularly Rhipicephalus sanguineus (the brown dog tick) in transmitting L. infantum has been investigated. In the present paper, the presence of Leishmania DNA was investigated in questing Ixodes ricinus ticks collected from 4 rural areas included in three parks of the Emilia-Romagna Region (north-eastern Italy), where active foci of human visceral leishmaniasis have been identified. The analyses were performed on 236 DNA extracts from 7 females, 6 males, 72 nymph pools, and 151 larvae pools. Four samples (1.7%) (i.e., one larva pool, 2 nymph pools, and one adult male) tested positive for Leishmania kDNA. To the best of our knowledge, this is the first report of the presence of Leishmania kDNA in questing I. ricinus ticks collected from a rural environment. This finding in unfed larvae, nymphs, and adult male ticks supports the hypothesis that L. infantum can have both transstadial and transovarial passage in I. ricinus ticks. The potential role of I. ricinus ticks in the sylvatic cycle of leishmaniasis should be further investigated. [ABSTRACT FROM AUTHOR]

Kala azar elimination programme in India is mainly depends on vector control measures and primary methods of application of insecticides is indoor residual spray (IRS). For vector control IRS should be carried out at regular intervals of six months. Presently, alphacypermethrin is used for IRS. This study was designed to study the impact of IRS with alphacypermethrin by analysing monthly population density following IRS-treatment. Twenty villages of Habibpur block, Malda district was selected for study which were grouped into study arm where IRS was done and control arm where IRS was not done. This study was conducted during April 2019 to June, 2020. In study villages, IRS was done during August, 2019 (1st cycle) and January 2020 (2nd cycle). Before and after IRS, entomological survey was carried out in cattle shades, human habitation and peri-domestic vegetations using mouth aspirators and light traps. Man hour density, per trap per night collection and % of reduction of sand fly abundance was calculated from survey result. In study arm after 14 days of 1st cycle of IRS, sand fly population reduced at rate of 96.5% which continues upto 3 months (85.34%). About 90% sand fly reduced after 14 days but after 3 months of 2nd cycle IRS density increased by 11.6%. Similar finding was recorded in the both cattle sheds and human habitations. Light trap collection from peri-domestic vegetation in both study and control villages were almost similar throughout the study period. Higher reduction rate of sand fly population following IRS showed that the used insecticide is effective against prevailing vector population of the study areas. The IRS has no effect on peri-domestic population. The presence of sand flies out side human habitation and cattle shades might play an important role in increasing population density following reduced residual activity of insecticide. [ABSTRACT FROM AUTHOR]

The purpose of the present study was to evaluate the efficacy of the treatment with a recombinant cysteine proteinase from Leishmania, rldccys1, associated with allopurinol or miltefosine on Leishmania (Leishmania) infantum chagasi–infected hamsters. Golden Syrian hamsters infected with L. (L.) infantum chagasi were treated with either miltefosine (46 mg/kg) or allopurinol (460 mg/kg) alone by oral route or associated with rldccys1 (150 µg/hamster) by subcutaneous route for 30 days. Infected hamsters were also treated with miltefosine (46 mg/kg) plus rldccys1 (150 µg/hamster) for 30 days (phase 1) followed by two additional doses of rldccys1 (250 µg/hamster) (phase 2). After the end of treatment, the animals were analyzed for parasite load, body weight, serum levels of immunoglobulins, cytokine expression, and drug toxicity. The data showed a significant decrease of parasite load in infected hamsters treated with allopurinol or miltefosine alone or associated with rldccys1, as well as in those treated with rldccys1 alone. Significantly lower levels of serum IgG were detected in hamsters treated with allopurinol plus rldccys1. The treatment with miltefosine associated with rldccys1 prevented relapse observed in animals treated with miltefosine alone. A significant loss of body weight was detected only in some hamsters treated with miltefosine for 1 month and deprived of this treatment for 15 days. There were no significant differences in transcript expression of IFN-γ and IL-10 in any of treated groups. Neither hepatotoxicity nor nephrotoxicity was observed among controls and treated groups. These findings open perspectives to further explore this immunochemotherapeutic schedule as an alternative for treatment of visceral leishmaniasis. [ABSTRACT FROM AUTHOR]

This study aimed to characterize agglutinating antibodies detected by the direct agglutination test (DAT-LPC) for the diagnosis of visceral leishmaniasis (VL). The DAT-LPC antigen/antibodies complex was recovered, washed, and used as antigenic substrate in a modified enzyme-linked immunosorbent assay (modified ELISA), revealed with anti-human IgM, IgG, and IgG subtype conjugates, and in the immunofluorescent antibodies test (IFAT), revealed with anti-human IgG and IgG1 conjugates. IgM antibodies were detected in 50%, IgG and IgG1 in 100%, and IgG3 in 52.8% of the 36 samples from VL patients. IFAT showed that agglutinating IgG and IgG1 antibodies recognized more intensely antigens located in the membrane and kinetoplast of the parasite. No antibodies were detected in the 15 samples from healthy individuals. This study shows for the first time that the antibodies responsible for agglutination in DAT-LPC are mostly of the IgG1 subtype. [ABSTRACT FROM AUTHOR]

The use of repellents is considered an alternative against biting insects, including Lutzomyia longipalpis (Diptera: Psychodidae), the main vector of the protozoan Leishmania infantum, visceral leishmaniasis's (VL) etiologic agent in the Americas. This study aimed to evaluate the repellent efficacy of icaridin nanostructured solution applied on cotton knitting fabric against L. longipalpis. Arm-in-cage tests were performed in eight volunteers at different concentrations (5%, 10%, 25%, and 50%), using L. longipalpis (n = 30). The bioassay was performed in 1, 24, 48, 72, 96, 120, and 144 h after impregnation and one test after washing the fabrics with icaridin. The total repellency rate (%R) > 95% was used as a reference to define a minimum effective concentration (MEC). The results revealed that the insects' landing mean decreased significantly in different icaridin concentrations, compared with the control tests (p < 0.05) and the 25% and 50% concentrations compared to lower concentration (5%) (p < 0.05). The higher concentrations (25% and 50%) provided longer complete protection times (CPTs) with 120 and 144 h of protection, respectively and the %R of 100% for 72 and 96 h after impregnation, respectively. The 25% was the MEC (%R Total = 98.18%). Our results indicate, for the first time, that icaridin nanostructured solution applied on cotton knitting fabric proved to be an efficient repellent against L. longipalpis with the presence of repellent action even after washing. The concentration of 25% showed better efficiency and may become an efficient method for L. longipalpis biting control. [ABSTRACT FROM AUTHOR]

The parasitic protozoa Leishmania (Leishmania) infantum is the etiological agent of human visceral leishmaniasis and canine leishmaniasis in South America, where Brazil is the most affected country. This zoonotic disease is transmitted by the bite of an infected phlebotomine sand fly and dogs constitute the main domestic reservoir of the parasite. In this study, we screened 2348 dogs of the municipality of Embu das Artes, Brazil, for antibodies against the parasite. Prevalence for canine leishmaniasis seropositivity was 2.81%, as assessed using a Dual-Path Platform rapid test for canine leishmaniasis. Twenty-five seropositive dogs were euthanized for parasite isolation and 14 isolates were successful obtained. Nucleotide sequencing of the internal transcribed spacer confirmed the isolates to be L. (L.) infantum, and very low sequence variability was observed among them. The in vitro susceptibility to miltefosine and paromomycin was assessed and moderate variation in paromomycin susceptibility was found among the isolates in the promastigote and intracellular amastigote stages. On the other hand, in vitro susceptibility to miltefosine of these isolates was homogenous, particularly in the amastigote stage (EC50 values from 0.69 to 2.07 μM). In addition, the miltefosine sensitivity locus was deleted in all the isolates, which does not corroborate the hypothesis that the absence of this locus is correlated with a low in vitro susceptibility. Our findings confirm that the municipality of Embu das Artes is endemic for canine leishmaniasis and that isolates from this region are susceptible to paromomycin and miltefosine, indicating the potential of these drugs to be clinically evaluated in the treatment of human visceral leishmaniasis in Brazil. [ABSTRACT FROM AUTHOR]

Purpose: Leishmaniasis is a disease caused by different Leishmania spp., which are transmitted to humans by a bite of infected female sand flies. Cutaneous leishmaniasis (CL, oriental sore), visceral leishmaniasis (VL), and mucocutaneous leishmaniasis (MCL) are three main clinical forms, however, only CL and VL are seen in Turkey. Cutaneous leishmaniasis is characterized by skin lesion(s) and is one of the most important vector-borne diseases in Turkey with over 2000 cases reported annually in 40 out of 81 provinces. The treatment is usually made invasively and painfully by intralesional injection of pentavalent antimony compounds. Non-invasive and innovative treatment methods are needed as aimed in this study. Methods: In the present study, one of the classical antileishmanial drugs, amphotericin B (AmB), encapsulated in liposomes was evaluated using non-invasive design based on chitosan, which is a nontoxic, biocompatible and biodegradable polymer. To avoid the invasive effect of conventional intralesional needle application, the drug was encapsulated in liposomes and incorporated into a chitosan gel for applying topically on the skin lesion. The efficacy of encapsulation of amphotericin B into liposomes and the drug release from liposomes were studied. The chitosan gel was evaluated for viscosity, flowability, appearance and pH. The efficacy of the drug embedded into chitosan gel, liposomal AmB alone and chitosan gel alone in four different concentrations was also tested using Leishmania spp. promastigotes in vitro. Results: The findings have shown that AmB was encapsulated into the liposomes with high efficiency (86.6%) and long-term physical and chemical stability. Therefore, designed liposomal formulation was suitable for sustained release. The appearance of the drug-embedded chitosan gel was transparent and appropriate. Chitosan gels showed non- Newtonian behavior and plastic flow. The liposomal AmB also showed higher efficacy with no parasites in all concentrations while drug embedded into chitosan gel and chitosan gel alone were effective in two higher concentrations. The lower efficacy of the drug-embedded chitosan gel in 24 h in in-vitro study was probably due to slow release of the drug. Conclusion: The gel design created in this study will provide ease of use for the lesions of CL patients that do not have a specific number, size, and shape. Follow-up studies by the ex-vivo macrophage infection model with Leishmania intracellular amastigote forms and Leishmania-infected animal models are needed to understand the present design's efficacy better. [ABSTRACT FROM AUTHOR]

Asymptomatic visceral leishmaniasis cases increase continuously, particularly among patients with HIV who are at risk to develop further symptoms of leishmaniasis. A simple, sensitive and reliable diagnosis is crucially needed due to risk populations mostly residing in rural communities with limited resources of laboratory equipment. In this study, a highly sensitive and selective determination of Leishmania among asymptomatic patients with Leishmania/HIV co-infection was achieved to simultaneously interpret and semi-quantify using colorimetric precipitates (gold-nanoparticle probe; AuNP-probe) and fluorescence (SYBR safe dye and distance-based paper device; dPAD) in one-step loop-mediated isothermal amplification (LAMP) assay. The sensitivities and specificities of 3 detection methods were equivalent and had reliable performances achieving as high as 95.5%. Detection limits were 102 parasites/mL (0.0147 ng/µL) which were 10 times more sensitive than other related studies. To empower leishmaniasis surveillance as well as prevention and control, this dPAD combined with SYBR safe and gold nanoparticle probe LAMP assay is reliably fast, simple, inexpensive and practical for field diagnostics to point-of-care settings in resource-limited areas which can be set up in all levels of healthcare facilities, especially in low to middle income countries. [ABSTRACT FROM AUTHOR]

Kala-azar/Visceral Leishmaniasis (VL) caused by Leishmania donovani (LD) is often associated with Leptomonas seymouri (LS) co-infection in India. Leptomonas seymouri narna-like virus 1 (Lepsey NLV1) has been reported in multi-passaged laboratory isolates of VL samples which showed LD-LS co-infection. A pertinent question was whether this virus of LS is detectable in direct clinical samples. DNA from the serum of twenty-eight LD diagnosed patients was subjected to LD-specific and LS-specific PCR to reconfirm the presence of LD parasites and to detect LD-LS co-infections. RNA extracted from same samples was subjected to RT-PCR, qRT-PCR and sequencing using virus-specific primers to detect/identify and quantify the virus. The presence of the virus was confirmed in thirteen of eighteen (72%) recently collected VL and PKDL samples. Cytokine profiling showed significantly elevated IL-18 in only LD infected patients compared to the virus-positive LD and control samples. IL-18 is crucial for Th1 and macrophage activation which eventually clears the parasite. The Lepsey NLV1 interaction with the immune system results in reduced IL-18 which favors LD survival and increased parasitic burden. The study emphasizes the need to revisit LD pathogenesis in the light of the association and persistence of a protozoan virus in kala-azar and PKDL patients. [ABSTRACT FROM AUTHOR]

Visceral leishmaniasis (VL) is a neglected tropical disease that mainly affects the poor population of the Indian, African, and South American subcontinent. The increasing resistance to antimonial and miltefosine and frequent toxicity of amphotericin B drives an urgent need to develop an anti-leishmanial drug with excellent efficacy and safety profile. In this study, three sequential docking protocols (HTVS, SP, and XP) were performed to screen the secondary metabolites (n = 6519) from the actinomycetes source against five key proteins involved in the metabolic pathway of Leishmania donovani. Those proteins were adenine phosphoribosyltransferase (PDB ID: 1QB7), trypanothione reductase (PDB ID: 2JK6), N-myristoyl transferase (PDB ID: 2WUU), pteridine reductase (PDB ID: 2XOX), and MAP kinase (PDB ID: 4QNY). Although the binding energy of top ligands was predicted using the MM-GBSA module of the Schrödinger suite. SP and XP docking mode resulted in 55 multi-targeted ligands against L donovani. MM-GBSA analysis selected the top 18 ligands with good-binding affinity and the binding-free energy for four proteins, as mentioned earlier, when compared with the miltefosine, paromomycin, and a reference ligand selected for each target. Finally, molecular dynamics simulation, post-MD-binding-free energy (MM-PBSA), and principal component analysis (PCA) proposed three best ligands (Adenosine pentaphosphate, Atetra P, and GDP-4-keto-6-deoxymannose) qualifying the above screening parameters and confirmed as a potential drug candidate to fight against Leishmania donovani parasites. [ABSTRACT FROM AUTHOR]